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Journal article

Imaging cell interaction in tracheal mucosa during influenza virus infection using two-photon intravital microscopy

  • Palomino-Segura, Miguel Institute for Research in Biomedicine (IRB), Faculty of Biomedical Sciences, Università della Svizzera italiana, Switzerland - Graduate School of Cellular and Molecular Sciences, Faculty of Medicine, University of Bern
  • Virgilio, Tommaso Institute for Research in Biomedicine (IRB), Faculty of Biomedical Sciences, Università della Svizzera italiana, Switzerland - Graduate School of Cellular and Molecular Sciences, Faculty of Medicine, University of Bern
  • Morone, Diego Institute for Research in Biomedicine (IRB), Faculty of Biomedical Sciences, Università della Svizzera italiana, Switzerland
  • Pizzagalli, Diego U. Institute for Research in Biomedicine (IRB), Faculty of Biomedical Sciences, Università della Svizzera italiana, Switzerland - Istituto di scienza computazionale (ICS), Facoltà di scienze informatiche, Università della Svizzera italiana, Svizzera
  • Gonzalez, Santiago F. Institute for Research in Biomedicine (IRB), Faculty of Biomedical Sciences, Università della Svizzera italiana, Switzerland
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    17.08.2018
Published in:
  • Journal of visualized experiments. - 2018, vol. 138, p. e58355
Immunology and infection
Issue 138
Two-photon intravital microscopy
Trachea
Influenza virus
Neutrophil
Dendritic cell
Cell-to-cell interactions
English The analysis of cell-cell or cell-pathogen interaction in vivo is an important tool to understand the dynamics of the immune response to infection. Two-photon intravital microscopy (2P-IVM) allows the observation of cell interactions in deep tissue in living animals, while minimizing the photobleaching generated during image acquisition. To date, different models for 2P-IVM of lymphoid and non-lymphoid organs have been described. However, imaging of respiratory organs remains a challenge due to the movement associated with the breathing cycle of the animal. Here, we describe a protocol to visualize in vivo immune cell interactions in the trachea of mice infected with influenza virus using 2P-IVM. To this purpose, we developed a custom imaging platform, which included the surgical exposure and intubation of the trachea, followed by the acquisition of dynamic images of neutrophils and dendritic cells (DC) in the mucosal epithelium. Additionally, we detailed the steps needed to perform influenza intranasal infection and flow cytometric analysis of immune cells in the trachea. Finally, we analyzed neutrophil and DC motility as well as their interactions during the course of a movie. This protocol allows for the generation of stable and bright 4D images necessary for the assessment of cell-cell interactions in the trachea.
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  • English
Classification
Medicine
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https://n2t.net/ark:/12658/srd1318893
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